CN113980839B - Delftia sp NLG11 for degrading tobacco nicotine and application thereof - Google Patents

Abstract

The invention discloses a dalfter bacterium (Delftia sp.) NLG11 for degrading tobacco nicotine and application thereof, wherein the strain is preserved in Guangdong province microorganism strain preservation center at 28 th 6 th 2021, and the preservation number is GDMCC NO:61747. the research of the invention shows that the Delftia NLG11 strain has good degradation capability to nicotine under different pH values and temperatures, the maximum degradation capability is 70.1%, the smoke concentration and strength can be reduced, the comfort of smoke is improved, and the formula applicability of tobacco leaves is improved. The invention provides a strain capable of degrading nicotine in tobacco, and has good application value in tobacco industry and environmental protection.

Description

Delftia sp NLG11 for degrading tobacco nicotine and application thereof

Technical Field

The invention relates to the technical field of agricultural microorganisms, and particularly relates to a dalfot bacterium NLG11 for degrading tobacco nicotine and application thereof.

Background

In recent years, the appearance quality of tobacco leaves in partial tobacco areas of China is close to the international level, but a certain gap exists in terms of the inherent quality, and one of the more common and more prominent problems is higher nicotine content. How to reduce the nicotine content in these tobacco leaves is a real problem faced by each cigarette industry.

Nicotine, also called nicotine (nicotine), is a unique and most important alkaloid in various tobacco plants, has extremely strong activity, and is a main component in tobacco alkaloid. Nicotine is one of important factors influencing the quality of tobacco leaves, and the excessive content of nicotine in the tobacco leaves can increase the spicy irritation of smoke and influence the smoking taste, and the tobacco leaves have poor safety and harm health. In the process of smoking tobacco, nicotine is nitrited to generate the special strong carcinogen N-nitrosamine in tobacco. Tobacco can produce high-concentration nicotine waste in the processing process, and the environment is harmed. Therefore, the proper and stable nicotine content in the tobacco leaves is very important for ensuring the cigarette quality and maintaining the health of consumers.

Currently, there are three main ways to reduce nicotine content in tobacco: (1) chemical route: the alkaloid in the tobacco leaves can be removed by the method of treating the tobacco leaves by rinsing the tobacco leaves with hot water, extracting with an organic solvent, extracting with gas, distilling with steam and the like. (2) agricultural planting path: mainly controls from the aspects of heredity, ecology, cultivation and the like. (3) biotechnological pathways: the microbe capable of degrading nicotine is separated from tobacco or soil by using a microbe and enzyme method, and the microbe is directly acted on tobacco leaves after being cultured or after an enzyme system is separated out, so that the nicotine content in the tobacco leaves is reduced, and the quality of the tobacco leaves is improved. Although a part of nicotine can be removed by a chemical method, the cost is high, harmful byproducts are more, some aroma components in the tobacco are lost, the appearance color and luster are obviously changed, and the original good quality of the tobacco is influenced. With the development of science and technology, the method for controlling the nicotine content in tobacco products and tobacco waste by using microorganisms is efficient and high in selectivity, can reduce the nicotine content in tobacco, does not destroy the original excellent quality of the tobacco leaves, can increase the fragrance of the tobacco leaves, and can reduce green miscellaneous gas.

In tobacco and tobacco-growing soils, some microorganisms have adapted to environments containing nicotine and are able to use nicotine as a carbon nitrogen source and energy source for growth. The nicotine-tolerant microorganisms isolated at present mainly comprise 2 major groups of yeast and bacteria, wherein the bacteria mainly belong to 2 genera of Arthrobacter and Pseudomonas. The Arthrobacter genus mainly includes: arthrobacter nicotinovorans (Arthrobacter nicotinovorans), arthrobacter oxydans (Arthrobacter oxidans), arthrobacter ureafaciens (Arthrobacter ureafaciens), arthrobacter nicotianae (Arthrobacter nicotinovorans), and the like. The pseudomonas is mainly characterized in that: pseudomonas aeruginosa (Pseudomonas convoxa), pseudomonas putida (Pseudomonas putida), and the like. The current research mainly aims at separating and identifying nicotine-tolerant microorganisms, and the research shows that tobacco leaves are treated by using Arthrobacter (Arthrobacter spp.) K7 and K3 strain bacterial suspensions, the nicotine content is reduced by 26.1-39.7%, the leaf aroma quality and the leaf aroma quantity of the tobacco leaves are both increased, the smoke is fine and smooth, the irritation is reduced, the strength is reduced, and the aftertaste is improved (Lei Liping, xia Zhenyuan, guo Rongjun, wu Yuping, cui Guomin, liao Dezhi. The nicotine reducing effect of the Arthrobacter on the tobacco leaves [ J ] tobacco science and technology, 2008 (03): 56-58.); the strain ZUTSKD capable of efficiently metabolizing nicotine is also separated from pseudomonas and can grow by taking nicotine as a unique carbon-nitrogen source and energy source, and the degraded nicotine is used for treating tobacco waste (Sun Kedan, zhu Chenjing, zhong Weihong, chen Jianmeng, she Zijuan, liu Peiji and Zhou Jiang. The separation identification and degradation characteristics of a nicotine efficient degrading strain [ J ]. Environmental science report, 2008 (07): 1294-1301.).

The microbe bacteria which can tolerate and degrade nicotine are mainly two of Arthrobacter and Pseudomonas, the initial nicotine degrading concentration of the microbe bacteria obtained by separation is low, and few reports are reported about the Nicotine degrading of the Pond-derived Delftia sp. Patent CN107841474B discloses the prevention and treatment application of Povida Delftia lacustris ZJU-B1 strain to rice false smut, and the application of the strain in tobacco research and screening to obtain a new strain capable of resisting high nicotine concentration and high degradation efficiency is not seen, so that the problem to be solved at present is solved, and more efficient excellent strain selection is provided for microbial degradation of nicotine.

Disclosure of Invention

The invention provides a new excellent strain capable of degrading tobacco nicotine, and the strain has good application value in tobacco industry and environmental protection.

The first purpose of the invention is to provide a Delftia sp NLG11 strain.

The second purpose of the invention is to provide the application of the dalfot bacterium NLG11 strain in degrading tobacco nicotine.

A third object of the present invention is to provide a method for reducing nicotine in tobacco.

A fourth object of the invention is to provide a formulation for degrading nicotine.

The above object of the present invention is achieved by the following technical solutions:

a dalf bacteria (Delftia sp.) NLG11 strain, which was deposited at 28 months 6 of 2021 with the GDMCC (GDMCC) of the guangdong province, and the accession number of the strain is GDMCC NO:61747.

specifically, the nucleotide sequence of the 16S rDNA of the strain is shown in SEQ ID NO. 1.

The invention separates and obtains Delftia viviparum NLG11 strain from the midgut of brown planthopper, inoculates the Delftia viviparum NLG11 strain in a basic culture medium MSM containing nicotine (1 g/L) with the inoculation amount of 5 percent to culture for 72h, the cell of the strain NLG11 continuously proliferates within 0 to 36h, the cell concentration is reduced within 36 to 72h, and the maximum OD is reduced within 72h of culture time ₆₀₀ = 0.998. The maximum degradation capacity of the strain NLG11 to nicotine within 72 hours reaches 70.1 percent. The NLG11 strain separated and identified by the invention is shown to have the degradation effect on nicotine.

Therefore, the invention provides the application of the dalfot bacterium NLG11 strain or the bacterial liquid thereof in degrading tobacco nicotine.

The invention also provides application of the dalfot bacterium NLG11 strain or bacterial liquid thereof in preparation of products for degrading nicotine.

The invention also provides a method for reducing the nicotine in the tobacco, which adopts the Delftia sp NLG11 strain or the bacterial liquid thereof to degrade the nicotine in the tobacco.

Preferably, the NLG11 strain can degrade nicotine at 30-40 ℃.

More preferably, the NLG11 strain is capable of degrading nicotine at 30 ℃.

Preferably, the NLG11 strain is capable of degrading nicotine at pH6.0 to 8.0.

More preferably, the NLG11 strain is capable of degrading nicotine at pH 7.0.

Preferably, the method employs a concentration of 1 × 10 ⁸ ～1×10 ¹⁰ cfu/mL bacterial liquid of the Delftia novyi NLG11 bacterial strain is sprayed on the tobacco.

The invention also provides a preparation for degrading nicotine, which contains the Delftia sp NLG11 strain or the bacterial liquid thereof.

Preferably, the bacterial liquid concentration of the nicotine degrading preparation is 1 x 10 ⁸ ～1×10 ¹⁰ cfu/mL

Further preferably, the bacterial liquid concentration of the preparation is 1 × 10 ⁹ CFU/mL。

The invention also provides application of the nicotine-degrading preparation in degrading nicotine.

Compared with the prior art, the invention has the following beneficial effects:

the invention provides a nicotine-reducing dalbergia sp NLG11 strain, which is preserved in Guangdong province microorganism strain preservation center in 2021, 6 and 28 days, wherein the preservation number is GDMCC NO:61747. the research of the invention shows that the Delftia delavayi NLG11 strain has better degradation effect on nicotine at the temperature of 30 ℃ and the pH value of 7.0, and the strain NLG11 can reduce the smoke concentration and strength, improve the aroma quality of smoke, improve the formula applicability of tobacco leaves and keep the original quality characteristics of the tobacco leaves. The NLG11 strain can be used as a bacterium of a novel microbial preparation tobacco, is used for degrading the nicotine content of the tobacco, and has a good application prospect.

Drawings

FIG. 1 is a isolate plate of strain NLG11.

FIG. 2 is a 16S rDNA phylogenetic tree of strain NLG11 (note: marked in each branch as: genBank sequence number + strain name).

FIG. 3 is a colony of strain NLG11 on nicotine medium.

FIG. 4 shows the nicotine-degrading ability of strain NLG11.

Detailed Description

The invention is further described with reference to the drawings and the following detailed description, which are not intended to limit the invention in any way. The reagents, methods and apparatus employed in the present invention are conventional in the art, except as otherwise indicated.

Unless otherwise indicated, reagents and materials used in the following examples are commercially available.

EXAMPLE 1 isolation of the strains

1. Preparation of selective separation culture medium

LB medium: 10g of peptone, 5g of yeast extract, 10g of NaCl,15g of QiongongLipid, dissolved in 1L sterile ddH ₂ And adjusting the pH to 7.0 in O water.

EB medium: 3g of beef extract powder, 10g of tryptone, 15g of yeast powder, 5g of sodium chloride, 10g of glucose and 15g of agar are dissolved in 1L of sterile water, and the pH value is adjusted to 7.0.

NA culture medium: 10g of tryptone, 3g of beef extract powder, 5g of sodium chloride and 15g of agar are dissolved in 1L of sterile water, and the pH value is adjusted to 7.0.

2. Bioassay and sampling

Selecting 15-day rice with uniform growth vigor, dividing the rice into two groups and repeating the rice three times, wherein each group repeats 4-6 rice. Adopting a rice seedling soaking method to carry out brown planthopper bioassay: pulling out the rice with roots, washing the rice with clear water, airing the rice in a shady and cool place until the surface is anhydrous, soaking the rice in a solution with the nicotine concentration of 160mg/L and the clear water for 1min, taking the rice out of the shady and cool place until the surface is anhydrous, wrapping the root of the rice with wet cotton, placing the rice in a raw measuring cup, inoculating 15 adult brown planthopper female adults for 1-2 days, placing the rice in an artificial climate box for culturing for 3 days, and then respectively sucking the groups of the living brown planthoppers. Climate box conditions: the temperature is 28 ℃; humidity is 85%; the photoperiod is 14-10 h.

3. Grinding and coating plate

The collected brown planthoppers are disinfected on the surfaces (70% alcohol soaking for 30s,2% sodium hypochlorite soaking for 30s, and shaking in clear water twice for 1min each time) and transferred into test tubes containing 200 mu L PBS, and then ground into homogenate by a hand-held electric grinder, and centrifuged slightly, and the supernatants are taken. Dilution of 3 concentration gradient, spread on three medium, each treatment is three repeat. Culturing in a 37 deg.C constant temperature incubator, and observing every 24 h.

4. Continuous purification culture

After single colonies grow in the selective culture medium, firstly selecting the single colonies according to the color, size and shape of the colonies, continuously streaking and purifying the single colonies on a corresponding culture medium for more than 5 times, then storing and photographing streak plates for separating strains, inoculating the single colonies into an LB liquid culture medium as shown in figure 1, shaking the strains at 220rpm to an exponential growth phase, storing glycerol bacteria, and freezing and storing in a refrigerator at-80 ℃ for later use.

EXAMPLE 2 identification of the strains

1. Morphological characteristics of bacterial colony

The colony is round, flat, yellow, opaque, smooth in surface and regular in edge after being placed on an LB plate and cultured for 24 hours at 37 ℃ in an incubator. The bacteria have facultative aerobe, the optimal growth temperature is 30 deg.c, the nutrient requirement is not high, and the bacteria grow well in common culture medium LB.

2. Physiological and biochemical characteristic determination

The results of the physiological and biochemical characteristics of the intestinal isolate are shown in table 1: can be used for rhamnose, melphalan, amygdalin, arabinose, beta-galactosidase, arginine bi-hydrolase, ornithine decarboxylation, citric acid, glucose, mannitol, sorbitol, and sucrose. Can not decompose H2S, urease, tryptophan deaminase, indole, 3-hydroxy butanone, gelatinase and lysine decarboxylase.

TABLE 1 physiological and biochemical characteristics of Strain NLG11

Note: "+" is positive reaction, and "-" is negative reaction.

3. 16S rDNA identification

The stored genomic DNA of the monoclonal strain was extracted using a bacterial genomic DNA extraction Kit (TIANAmp Bacteria DNA Kit) from Tiangen organisms, and PCR was performed using the extracted DNA as a template, 16S rDNA universal primers 27F (5 '-AGTTTGATCMTGGCTCAG-3') and 1492R (5'-GGTTACCTTGTTACGACTT-3') as upstream and downstream primers to amplify 16S rDNA of Bacteria, and the PCR reaction system is shown in Table 2. After mixing gently, centrifuging briefly, placing on a PCR instrument, and performing amplification procedures: pre-denaturation at 98 ℃ for 2min; denaturation at 98 ℃ for 10s, annealing at 55 ℃ for 15s, extension at 72 ℃ for 50s, and 30 cycles; 72 ℃ for 5min;10 ℃. The PCR product is detected by 1 percent agarose gel, cut, recovered and purified and then sent to Guangzhou Ongke biotechnology Limited company for sequencing and identification.

TABLE 2 1696 rDNA PCR amplification System

4. Construction of phylogenetic trees

The amplified product genome extracted in step 3 was subjected to sequence analysis using universal primers 27F and 1492R of bacterium 16S, and the sequence was spliced with SeqMan (DNAStar), and then blast-aligned with rRNA/ITS database in NCBI. Selecting a near sequence of a downloaded and finished intestinal isolate Delftia NLG11 strain, then performing multi-sequence comparison analysis by ClustalW software, then constructing a phylogenetic tree by Mega7.03 software by adopting an adjacency method (Neighbor-Joining), adjusting a bootstrap value and checking the reliability of the phylogenetic tree.

The results are shown in FIG. 2, and compared with Delftia lactisteristrin 3 (NR 116495.1. The strain is named as Delftia sp NLG11 strain, which is deposited in 28 days 6.2021 in Guangdong province microbial culture collection center with the deposit number of GDMCC NO:61747, deposited as Miehu Middy road No. 100, guangzhou, guangdong province.

Example 3 screening of Strain NLG11 in Nicotine Medium

Inorganic salt basal medium (MSM): 13.3g K ₂ HPO ₄ ，4.0g KH ₂ PO ₄ 0.1 g(NH ₄ ) ₂ SO ₄ Yeast powder 1.0g, trace elements 10.0mL (1.0 g MgSO) ₄ ·7H ₂ O，0.4g MnSO ₄ ·H ₂ O，0.2g CaCl ₂ ·2H ₂ O，0.2g CuCl ₂ ·2H ₂ O，0.02gFeSO ₄ ·7H ₂ O, dissolved in 100mL of 0.1mol/L HCl), and distilled water was added to make the volume 1L.

Nicotine culture medium: 90% nicotine was filtered through a 0.22 μm filter and added to the sterilized inorganic salt basal medium.

The activated NLG11 strain is scratched on MSM solid culture medium added with 200mg/L nicotine, and is inversely cultured for 24h at 30 ℃ in an incubator, and the growth of colonies in a culture dish is observed, and the result is shown in figure 3. As can be seen from FIG. 3, the NLG11 strain can grow single colony in 200mg/L MSM solid added with nicotine, and the colony is transparent.

Example 4 degradation Rate of Nicotine vs growth Rate of Strain NLG11

1. Drawing a nicotine standard curve

Spectrophotometric method: preparing the pure nicotine into working solution with the mass concentration of 5.0g/L by using 0.05mol/L HCl solution, respectively taking 0.01 mL, 0.02 mL, 0.03 mL, 0.04 mL, 0.05 mL and 0.06mL of the working solution, and fixing the volume to 10mL by using 0.05mol/LHCl solution. The absorbance at 259nm of nicotine solutions of different mass concentrations was determined with reference to 0.05mol/L HCl solution. When the mass concentration of the nicotine is measured to be 0.005-0.050 g/L by using a spectrophotometer method, the light absorption value (Y) is in positive correlation with the nicotine concentration (X), the regression equation is Y =36.457X +0.0053 ² Is 0.9997.

2. Preparation of NLG11 seed bacteria

The strain NLG11 glycerol, preserved at-80 ℃, was inoculated with 1:100 inoculating fresh liquid LB culture medium, activating at 220rpm and 37 ℃ for 14h to obtain seed bacteria, and then adding 1:100 of the culture medium, inoculating the strain in a fresh liquid LB culture medium, and performing shaking culture at 220rpm and 37 ℃ to logarithmic phase. And (3) centrifuging the bacterial liquid at 12000rpm for 5min to collect thalli, resuspending the thalli by using a sterilized basic salt culture medium MSM with the same volume, centrifuging the thalli, washing the thalli twice, and then resuspending the thalli by using a basic salt culture medium MSM to obtain the degraded seed bacteria NLG11.

3. Detection of initial nicotine degradation of strain NLG11

Activated degraded seed bacterium NLG11 was inoculated at 5% inoculum size in nicotine-containing (1 g/L) basal medium MSM, cultured at pH =7.0, 30 ℃,180rpm for 72h, and a non-inoculated nicotine medium was used as a control. Sampling at 6h, 12h, 24h, 36h, 48h and 72h, and measuring OD by spectrophotometry ₆₀₀ Absorbance of strain NLG11 at nm wavelength. Simultaneously sucking culture solution for 6h, 12h, 24h, 36h, 48h and 72h, centrifuging at 12000rpm at 4 ℃ for 10min, taking supernatant, filtering with 0.22 mu m aqueous phase filter membrane, and diluting the supernatant with HCl with concentration of 0.05mol/L to a proper light absorption value range. The change of the characteristic absorption peak of nicotine at 260nm is measured by using HCl with the concentration of 0.05mol/L as a reference solution.

As shown in FIG. 4, the cells of the strain NLG11 continued to proliferate for 0 to 36 hours, the cell concentration decreased for 36 to 72 hours, and the culture time was 72 hoursThe highest concentration is about 10 ⁹ CFU/mL; the nicotine concentration in the culture medium is obviously reduced after 24 hours, and the nicotine concentration curve tends to be flat at 48 hours. The maximum degradation capacity of strain NLG11 on nicotine is 70.1%.

4. Effect of different temperatures on growth of Strain NLG11 and on Nicotine degradation

The activated degradation seed bacterium NLG11 is inoculated in a basic culture medium MSM containing nicotine (1 g/L) according to the inoculation amount of 5 percent, and is cultured for 24 hours under the conditions of 25 ℃,30 ℃, 35 ℃ and 40 ℃ and 180rpm respectively, and the growth condition of the strain NLG11 is determined.

As shown in Table 3, the strain NLG11 grew at 25 ℃ to 40 ℃ and grew well at 30 ℃ to 40 ℃. The optimum growth temperature of the strain NLG11 is 30 ℃, and at the temperature, the strain NLG11 achieves the maximum growth amount and the highest nicotine degradation rate.

5. Effect of pH on growth of Strain NLG11 and on Nicotine degradation

Inoculating activated degraded seed bacteria NLG11 into a basic culture medium containing nicotine (1 g/L) according to the inoculation amount of 5%, culturing for 24h under the condition of 180rpm, performing shake culture for 36h by using culture media with different initial pH values, and determining the growth of cells and the nicotine degradation condition. As a result, as shown in Table 3, the strain NLG11 grew well at pH6.0 to 8.0. The optimum pH value for cell growth is 7.0, under the condition of the pH value, the strain NLG11 achieves the maximum growth amount, and the degradation rate of nicotine is also the highest.

TABLE 3 degradation rate of Nicotine by Strain NLG11 liquid Medium at different temperatures and pH

Example 5 sensory evaluation of tobacco treated with Strain NLG11

1. Strain NLG11 treated tobacco

After the activated degraded seed bacterium NLG11 is cultured to a logarithmic growth phase, the bacterium liquid is centrifuged at 12000rpm for 5min to collect the bacterium, the bacterium is resuspended by sterilized PBS with the same volume, centrifuged and washed twice. Is configured to have a concentration of 1 × 10 ⁹ CFU/mL. For Yunnan producing area of 2019 yearsThe specific method for treating the redried tobacco of grade B2F comprises the following steps: the prepared concentration is 1 multiplied by 10 ⁹ Uniformly spraying CFU/mL bacterial strain NLG11 bacterial liquid on redried tobacco strips, turning over the tobacco leaves gently, balancing water and standing for 30min; after the moisture is balanced, the tobacco leaves are placed in a shady and cool place indoors and are shaded for 3 days. After finishing, the tobacco leaves are cut into threads and dried, the moisture of the tobacco leaves is controlled to be 12% -12.5%, sensory quality evaluation is carried out, and the redried tobacco sprayed with PBS with the same volume and the samples of the same batch, which are not processed, of the grade B2F is used as a reference.

Wherein sensory quality score = (aroma quality × 0.35+ aroma amount × 0.25+ miscellaneous gas × 0.1+ irritation × 0.15+ aftertaste × 0.15) × 11.11.

2. Strain NLG11 treated tobacco sensory quality score

The bacterial strain NLG11 is used for treating B2F-grade redried cigarettes, and the measurement result is shown in Table 4, so that 2019 Yunnan B2F style characteristic indexes can be obviously improved, the smoke concentration and the strength are reduced, the comfort of smoke is improved, and the formula applicability of tobacco leaves is improved. The re-drying tobacco processing of the B2F grade has no obvious influence on the quality characteristic indexes of 2019 Yunnan B2F, and the original quality characteristics of tobacco leaves can be kept.

TABLE 4 results of redrying tobacco grade B2F treated with Strain NLG11

In conclusion, the screened Delftia delavayi NLG11 strain can degrade nicotine well, after B2F flue-cured tobacco in Yunnan producing areas is treated by the strain NLG11, the aroma quality of tobacco leaves can be improved, the concentration and the strength are reduced, the evaluation value in sensory evaluation is high, the nicotine content in the tobacco can be reduced, the original good quality of the tobacco leaves is not damaged, and the aroma of the tobacco leaves can be increased. The invention provides excellent strains for degrading nicotine for production capacity, and has good application value in tobacco industry and environmental protection.

The above embodiments are only intended to illustrate the technical solutions of the present invention and not to limit the scope of the present invention, and it is obvious to those skilled in the art that other variations or modifications can be made based on the above description and ideas, and all embodiments need not be exhaustive. Any modification, equivalent replacement, and improvement made within the spirit and principle of the present invention should be included in the protection scope of the claims of the present invention.

Sequence listing

<120> dalfot bacterium NLG11 for degrading tobacco nicotine and application thereof

<160> 1

<170> SIPOSequenceListing 1.0

<210> 1

<211> 1400

<212> DNA

<213> Delftia sp NLG11 Strain

<400> 1

atgcagtcga acggtaacag gtcttcggac gctgacgagt ggcgaacggg tgagtaatac 60

atcggaacgt gcccagtcgt gggggataac tactcgaaag agtagctaat accgcatacg 120

atctgaggat gaaagcgggg gaccttcggg cctcgcgcga ttggagcggc cgatggcaga 180

ttaggtagtt ggtgggataa aagcttacca agccgacgat ctgtagctgg tctgagagga 240

cgaccagcca cactgggact gagacacggc ccagactcct acgggaggca gcagtgggga 300

attttggaca atgggcgaaa gcctgatcca gcaatgccgc gtgcaggatg aaggccttcg 360

ggttgtaaac tgcttttgta cggaacgaaa aagctccttc taatacaggg ggcccatgac 420

ggtaccgtaa gaataagcac cggctaacta cgtgccagca gccgcggtaa tacgtagggt 480

gcgagcgtta atcggaatta ctgggcgtaa agcgtgcgca ggcggttatg taagacagat 540

gtgaaatccc cgggctcaac ctgggaactg catttgtgac tgcatggcta gagtacggta 600

gagggggatg gaattccgcg tgtagcagtg aaatgcgtag atatgcggag gaacaccgat 660

ggcgaaggca atcccctgga cctgtactga cgctcatgca cgaaagcgtg gggagcaaac 720

aggattagat accctggtag tccacgccct aaacgatgtc aactggttgt tgggaattag 780

ttttctcagt aacgaagcta acgcgtgaag ttgaccgcct ggggagtacg gccgcaaggt 840

tgaaactcaa aggaattgac ggggacccgc acaagcggtg gatgatgtgg tttaattcga 900

tgcaacgcga aaaaccttac ccacctttga catggcagga agtttccaga gatggattcg 960

tgctcgaaag agaacctgca cacaggtgct gcatggctgt cgtcagctcg tgtcgtgaga 1020

tgttgggtta agtcccgcaa cgagcgcaac ccttgtcatt agttgctaca ttcagttgag 1080

cactctaatg agactgccgg tgacaaaccg gaggaaggtg gggatgacgt caagtcctca 1140

tggcccttat aggtggggct acacacgtca tacaatggct ggtacagagg gttgccaacc 1200

cgcgaggggg agctaatccc ataaaaccag tcgtagtccg gatcgcagtc tgcaactcga 1260

ctgcgtgaag tcggaatcgc tagtaatcgc ggatcagcat gccgcggtga atacgttccc 1320

gggtcttgta cacaccgccc gtcacaccat gggagcgggt ctcgccagaa gtaggtagcc 1380

taaccgcaag gagggcgctc 1400

Claims (9)

1. A dalf bacteria (Delftia sp.) NLG11 strain, which is deposited in the Guangdong province culture collection center (GDMCC) at 28 th 6 th 2021, and the strain number is GDMCC NO:61747.

2. the use of the dalfot bacterium NLG11 strain or its bacterial liquid of claim 1 for degrading nicotine in tobacco.

3. The use of the dalfot bacterium NLG11 strain or its bacterial liquid of claim 1 in the preparation of products for degrading nicotine.

4. A method for reducing nicotine in tobacco, characterized in that nicotine in tobacco is degraded by using the bacterium Delftia NLG11 of claim 1 or a bacterial solution thereof.

5. The method of claim 4, wherein the degradation conditions are: the temperature is 25-40 ℃, and the pH value is 6.0-8.0.

6. The method of claim 4, wherein the concentration is 1 x 10 ⁸ ～1×10 ¹⁰ cfu/mL of a bacterial liquid of the Delftia sp NLG11 strain disclosed by claim 1 is sprayed on tobacco.

7. A nicotine-degrading agent comprising the Delftia sp NLG11 strain according to claim 1 or a bacterial solution thereof.

8. The preparation of claim 7, wherein the bacterial fluid concentration is 1 x 10 ⁸ ～1×10 ¹⁰ cfu/mL。

9. Use of a formulation according to claim 7 or 8 for degrading nicotine.

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